Larval survival and development of susceptible and resistant Ostrinia nubilalis (Lepidoptera: Pyralidae) on diet containing Bacillus thuringiensis

Abstract 1 Larval survival and development of Dipel‐susceptible and ‐resistant strains of European corn borer, Ostrinia nubilalis (Hübner), were assayed using diets incorporating low doses of a commercial formulation of Bacillus thuringiensis var. kurstaki Berliner (Dipel ES). 2 Larval mortality, growth and development, pupation rate and pupal weight were not significantly different between Dipel‐susceptible and ‐resistant strains when larvae were reared on a nontoxic control diet. 3 Larval mortality of Dipel‐resistant larvae did not significantly change as Dipel concentration increased at the tested concentrations, whereas mortality of Dipel‐susceptible larvae increased dramatically as Dipel concentration increased. 4 Larval development was significantly delayed when larvae were fed diets containing low doses of Dipel. 5 Pupation rate and pupal body weight declined as Dipel concentration increased but it decreased faster for the susceptible strain than for the resistant strain.     

Interactions between Nosema pyrausta (Microsporidia: Nosematidae) and Bacillus thuringiensis subsp. kurstaki in the European corn borer (Lepidoptera: Pyralidae).

Larval susceptibility to Bacillus thuringiensis was determined for Nosema pyrausta-infected and uninfected European corn borers, Ostrinia nubilalis (Hübner), in bioassays using a commercial formulation of B. thuringiensis subsp. kurstaki, Dipel ES, incorporated into diet. LC50 values for N. pyrausta-infected larvae were significantly lower (P<0.0001) than for uninfected larvae and declined with increasing levels of infection. LC50 values for a 15-d bioassay using field-colony first instars were 0.006 and 0.027 mg of Dipel ES/kg of diet for larvae moderately infected by N. pyrausta and uninfected larvae, respectively. Nosema pyrausta-infected larvae reared on Dipel ES-amended diets produced 70-fold fewer spores (P<0.0001) than larvae reared on standard diet. For example, 15 d after placement as first instars on standard diet, infected field-colony larvae produced 7.6-8.7 million N. pyrausta spores per larva; similar larvae placed on diet containing 0.09 mg of Dipel ES/kg of diet produced 85-103 thousand spores per larva. Infected larvae also weighed less and failed to mature on Dipel ES-amended diets. Increased susceptibility of N. pyrausta-infected larvae to Dipel ES and reduced N. pyrausta spore production in larvae feeding on diet containing Dipel ES suggest that Bt corn will have a direct adverse effect on the survival and continual impact of N, pyrausta as a regulating factor on European corn borer populations.     

Host suitability and fitness-related parameters of Campoletis sonorensis (Hymenoptera: Ichneumonidae) as a parasitoid of the cabbage looper,Trichoplusia ni (Lepidoptera: Noctuidae)

The seven age-classes of Trichoplusia ni (Hübner) larvae evaluated in this study as hosts of Campoletis sonorensis indicates that early 2nd larval instar (3–5 day-old larvae) of T. ni represents the most suitable host stage for the development of the larval endoparasitoid C. sonorensis. The higher suitability of early 2nd larval instar of T. ni resulted in more parasitised larvae, a higher rate of successful parasitoid emergence, a higher rate of female progeny, and a lower rate of immature parasitoid mortality. The fitness gain of C. sonorensis on late 1st larval instar (2 day-old larvae) and late 2nd larval instar –early 3rd instars (6–8 day-old larvae) stages of T. ni is negatively affected by the trade-offs between the different physiological and behavioral characteristics influencing their suitability as hosts of C. sonorensis.     

Inheritance of resistance to Bacillus thuringiensis Cry1Ac toxin in a greenhouse-derived strain of cabbage looper (Lepidoptera: Noctuidae)

A population of cabbage looper, Trichoplusia ni (Hübner), collected from commercial greenhouses in the lower mainland of British Columbia, Canada, in 2001 showed a resistance level of 24-fold to Dipel, a product of Bacillus thuringiensis (Bt) subspecies kurstaki. This population was selected with Cry1Ac, the major Bt Cry toxin in Dipel, to obtain a homogenous population resistant to Cry1Ac. The resulting strain of T. ni, named GLEN-Cry1Ac, was highly resistant to Cry1Ac with a resistance ratio of approximately 1000-fold. The larvae from the GLEN-Cry1Ac strain could survive on Cry1Ac-expressing transgenic broccoli plants that were highly insecticidal to T. ni and diamondback moth, Plutella xylostella (L.). The inheritance of Cry1Ac resistance in this T. ni strain was autosomal and incompletely recessive. The degree of dominance of the resistance was -0.402 and -0.395, respectively, for the neonates in reciprocal crosses between the GLEN-Cry1Ac and a laboratory strain of T. ni. Using chi2 goodness-of-fit test, we demonstrated that the inhibition of larval growth resulting from testing 12 toxin doses in the progeny of the backcross fit the predicted larval responses based on a monogenic inheritance model. Therefore, we conclude that the inheritance of the resistance to Cry1Ac in the T. ni larvae is monogenic.     

Resistance to Bacillus thuringiensis Toxin Cry2Ab in Trichoplusia ni Is Conferred by a Novel Genetic Mechanism

The resistance to the Bacillus thuringiensis (Bt) toxin Cry2Ab in a greenhouse-originated Trichoplusia ni strain resistant to both Bt toxins Cry1Ac and Cry2Ab was characterized. Biological assays determined that the Cry2Ab resistance in the T. ni strain was a monogenic recessive trait independent of Cry1Ac resistance, and there existed no significant cross-resistance between Cry1Ac and Cry2Ab in T. ni. From the dual-toxin-resistant T. ni strain, a strain resistant to Cry2Ab only was isolated, and the Cry2Ab resistance trait was introgressed into a susceptible laboratory strain to facilitate comparative analysis of the Cry2Ab resistance with the susceptible T. ni strain. Results from biochemical analysis showed no significant difference between the Cry2Ab-resistant and -susceptible T. ni larvae in midgut proteases, including caseinolytic proteolytic activity and zymogram profile and serine protease activities, in midgut aminopeptidase and alkaline phosphatase activity, and in midgut esterases and hemolymph plasma melanization activity. For analysis of genetic linkage of Cry2Ab resistance with potential Cry toxin receptor genes, molecular markers for the midgut cadherin, alkaline phosphatase (ALP), and aminopeptidase N (APN) genes were identified between the original greenhouse-derived dual-toxin-resistant and the susceptible laboratory T. ni strains. Genetic linkage analysis showed that the Cry2Ab resistance in T. ni was not genetically associated with the midgut genes coding for the cadherin, ALP, and 6 APNs (APN1 to APN6) nor associated with the ABC transporter gene ABCC2. Therefore, the Cry2Ab resistance in T. ni is conferred by a novel but unknown genetic mechanism.     

Bacillus thuringiensis plants expressing Cry1Ac,Cry2Ab and Cry1F are not toxic to the assassin bug,Zelus renardii

Cotton‐ and maize‐producing insecticidal crystal (Cry) proteins from the bacterium, Bacillus thuringiensis (Bt), have been commercialized since 1996. Bt plants are subjected to environmental risk assessments for non‐target organisms, including natural enemies that suppress pest populations. Here, we used Cry1F‐resistant Spodoptera frugiperda (J.E. Smith) and Cry1Ac and Cry2Ab‐resistant Trichoplusia ni (Hübner) as prey for the assassin bug, Zelus renardii (Kolenati), a common predator in maize and cotton fields. In tritrophic studies, we assessed several fitness parameters of Z. renardii when it fed on resistant S. frugiperda that had fed on Bt maize expressing Cry1F or on resistant T. ni that had fed on Bt cotton expressing Cry1Ac and Cry2Ab. Survival, nymphal duration, adult weight, adult longevity and female fecundity of Z. renardii were not different when they were fed resistant‐prey larvae (S. frugiperda or T. ni) reared on either a Bt crop or respective non‐Bt crops. ELISA tests demonstrated that the Cry proteins were present in the plant at the highest levels, at lower levels in the prey and at the lowest levels in the predator. While Z. renardii was exposed to Cry1F and Cry1Ac and Cry2Ab when it fed on hosts that consumed Bt‐transgenic plants, the proteins did not affect important fitness parameters in this common and important predator.     

甜菜夜蛾羧酸酯酶基因cDNA的克隆、表达及序列分析

羧酸酯酶是昆虫体内重要的解毒酶系之一,与昆虫抗药性产生相关。利用粉纹夜蛾Trichoplusia ni (Hübner)肠粘蛋白多克隆抗体免疫筛选甜菜夜蛾Spodoptera exigua (Hübner)中肠cDNA表达文库,得到编码羧酸酯酶的全长cDNA克隆。该cDNA克隆全长1 812 bp(GenBank登录号EF580101),开放阅读框长1 605 bp,编码535个氨基酸。该蛋白活性中心包括3个氨基酸残基（催化三联体）： Ser186, Glu319和His443, 3个N-联糖基化位点,具备羧酸酯酶的结构特征,属羧酸酯酶家族（EC: 3.1.1.-）。将该基因与pQE30载体重组,经IPTG诱导,spot-blot鉴定,蛋白获得了表达;以α-醋酸萘酯为底物,检测表达的羧酸酯酶活性为1.3 nmol/100 μL酶液。     

Assessment of microencapsulated formulations for improved residual activity of Bacillus thuringiensis

Bacillus thuringiensis Berliner is a highly efficacious bioinsecticide used to control lepidopteran pests in the field. Unfortunately, it has limited residual activity on plants because sunlight inactivates spores and crystals and they can be washed off by rain. To minimize loss of activity, formulations must contain UV protectants, stickers, or both. We tested approximately 80 formulations and determined optimal combinations of ingredients and spray drying conditions for improving B. thuringiensis residual activity after simulated rain and simulated sunlight. B. thuringiensis stability, after simulated sunlight (xenon light/8 h) and rain (5 cm/50 min), was improved using formulations based on lignin, corn flours, or both, with up to 20% of the active ingredient, when compared with technical powder or Dipel 2x in laboratory assays. Two formulations, made with corn flours or lignin + pregelatinized corn flour (PCF), killed 51.6 and 75.3% of Ostrinia nubilalis (Hübner) neonates after rain, respectively, versus 27% for technical powder. When the insecticidal activity was tested after simulated sunlight, corn flour-based formulations killed 78.5% of test larvae, and the lignin + PCF formulation killed 70.4%, in contrast to technical powder which caused an average of 29% mortality. Formulations made with Dipel 2x rather than technical powder, caused 62.5% mortality (corn flour-based formulations), and 72.3% mortality (lignin + PCF), versus 53.4% for Dipel 2x after rain. When tested after simulated sunlight, formulations killed 95% of the larvae (average of both formulations) versus 82% for Dipel 2x. In a field test, formulations were applied to cabbage and insecticidal activity was determined against Trichoplusia ni (Hübner) neonates exposed to treated leaves. Insecticidal activity of the corn flour-based formulations was comparable to Dipel 2x for 4 d after treatment, but was significantly better than Dipel 2x 7 d after application. A lignin and PCF-based formulation showed significantly higher residual activity than Dipel 2x, 4 and 7 d after application.     

Development and encapsulation of the endoparasitoid,Microplitis croceipes (Hym.: Braconidae), in six candidate host species (Lep.)

Encapsulation and development of the endoparasitoid,Microplitis croceipes (Cresson), were studied in six atypical lepidopteran host species whose usual host isHelicoverpa zea (Boddie). The candidate hosts examined were: the fall armywormSpodoptera frugiperda (J. E. Smith); the beet armyworm,Spodoptera exigua (Hübner); the cabbage looper,Trichoplusia ni (Hübner); the greater wax moth,Galleria mellonella (L.); the Indian meal moth,Plodia interpunctella (Hübner); and the diamondback moth,Plutella xylostella (L.). BothS. exigua andT. ni were completely unsuitable forM. croceipes development due to the high rate of eggs that were encapsulated within three days after parasitism. Encapsulation inS. frugiperda included mainly parasitoid eggs and was first detected six days after parasitization at 25°C and two days at 30°C. Encapsulation inG. mellonella occurred only in the larval stage of the parasitoid. InP. interpunctella, parasitoid larvae reached the 3rd stadium, but none of them pupated. OnlyS. frugiperda andG. mellonella supported successful development ofM. croceipes from egg to adult. The percentage of parasitoids reaching the adult stage in these hosts was higher at 30°C than at 25°C (13% vs. 4% inS. frugiperda, and 21% vs. 3% inG. mellonella, respectively). However, these percentages were too low to substitute them as a more economical host for rearingM. croceipes. This biological information will be useful in additional laboratory studies directed toward reducing the rate of encapsulation (e.g., manipulation of host rearing temperature) to increase production ofM. croceipes on these hosts.     

Host plant species affects the quality of the generalist Trichoplusia ni as a host for the polyembryonic parasitoid Copidosoma floridanum

Diet of herbivorous insects can influence both the herbivores and their natural enemies. We examined the direct and indirect effects of diet on the interactions between the polyphagous herbivore Trichoplusia ni Hübner (Lepidoptera: Noctuidae) and its polyembryonic parasitoid Copidosoma floridanum Ashmead (Hymenoptera: Encyrtidae). To determine how host plant species and host plant iridoid glycoside content affect host caterpillars and their parasitoids, parasitized and unparasitized T. ni were given leaves of either Plantago lanceolata L., which contains the iridoid glycosides aucubin and catalpol, Plantago major L. (Plantaginaceae), which contains only aucubin, or Taraxacum officinale F.H. Wigg (Asteraceae), which contains neither. Survival of unparasitized T. ni was much lower when fed P. major compared with the other two host plants, whereas pupae were smallest when fed T. officinale and developed most slowly when fed P. lanceolata as larvae. Neither aucubin nor catalpol were detected in intact Plantago‐fed T. ni larvae or their hemolymph, and only trace amounts of aucubin were detected in frass, suggesting that these compounds are mostly metabolized in the midgut and are not encountered by the parasitoid. Copidosoma floridanum clutch size was almost doubled when reared from P. lanceolata‐fed T. ni compared with T. officinale‐fed larvae and tripled compared with P. major‐fed larvae, although the percent of parasitoids surviving to adulthood was uniformly high regardless of host diet. The observed variation in C. floridanum fitness among host diets is likely mediated by the effect of the diets on host quality, which in turn may be influenced more by other factors in the host plants than their iridoid glycoside profiles. Interactions between plant metabolites, generalist herbivores like T. ni, and their parasitoids may be predominantly indirect.     

Bacillus thuringiensis Bel Protein Enhances the Toxicity of Cry1Ac Protein to Helicoverpa armigera Larvae by Degrading Insect Intestinal Mucin

Bacillus thuringiensis has been used as a bioinsecticide to control agricultural insects. Bacillus cereus group genomes were found to have a Bacillus enhancin-like (bel) gene, encoding a peptide with 20 to 30% identity to viral enhancin protein, which can enhance viral infection by degradation of the peritrophic matrix (PM) of the insect midgut. In this study, the bel gene was found to have an activity similar to that of the viral enhancin gene. A bel knockout mutant was constructed by using a plasmid-free B. thuringiensis derivative, BMB171. The 50% lethal concentrations of this mutant plus the cry1Ac insecticidal protein gene were about 5.8-fold higher than those of the BMB171 strain. When purified Bel was mixed with the Cry1Ac protein and fed to Helicoverpa armigera larvae, 3 μg/ml Cry1Ac alone induced 34.2% mortality. Meanwhile, the mortality rate rose to 74.4% when the same amount of Cry1Ac was mixed with 0.8 μg/ml of Bel. Microscopic observation showed a significant disruption detected on the midgut PM of H. armigera larvae after they were fed Bel. In vitro degradation assays showed that Bel digested the intestinal mucin (IIM) of Trichoplusia ni and H. armigera larvae to various degrading products, similar to findings for viral enhancin. These results imply Bel toxicity enhancement depends on the destruction of midgut PM and IIM, similar to the case with viral enhancin. This discovery showed that Bel has the potential to enhance insecticidal activity of B. thuringiensis-based biopesticides and transgenic crops.Bacillus thuringiensis is a ubiquitous gram-positive, spore-forming soil bacterium and produces insecticidal crystal proteins during the sporulation phase of its growth cycle. Because these insecticidal crystal proteins have activity against certain insect species, B. thuringiensis has been extensively used as a biopesticide to control crop pests in commercial agriculture and forest management. It is also a key source of genes for transgenic expression and provides pest resistance in plants (2, 20, 30).The viral enhancin protein was originally described for granuloviruses (GVs) as a 126-kDa protein that showed an ability to enhance the infectivity of nucleopolyhedroviruses (NPVs) (36, 37, 39). It has also been found in several other GVs (13) and NPVs (19, 27). Considered a pathogenicity factor, it is not essential for growth of viruses in cell culture or infected insects but has the function of facilitating GV and NPV infection and decreasing larval survival time (14, 17, 19, 27).The widely accepted action mode of the viral enhancin protein, which has been identified as a metalloprotease (17), is that it can disrupt the protective peritrophic matrix (PM), allowing virion access to the underlying epithelial cells of the insect gut (17). The PM has a lattice structure formed by chitin and insect intestinal mucin (IIM), and the viral enhancin protein targets the IIM for degradation (33).Enhancin-like genes with 24 to 25% nucleotide identity to viral enhancin genes have been found in Yersinia pestis, Bacillus anthracis, Bacillus thuringiensis, and Bacillus cereus genome sequences (16, 25, 28). When B. cereus enhancin-like protein was expressed in recombinant Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV) budded viruses and polyhedral inclusion bodies, it was found to be cytotoxic compared to viral enhancin protein. However, larval bioassays indicated that this enhancin-like protein did not enhance infection (8). Hajaij-Ellouze et al. (12) isolated a B. thuringiensis enhancin-like gene from a 407 crystal-minus strain and found that this enhancin-like protein has a typical metalloprotease zinc-binding domain (HEIAH) and belongs to the PlcR regulon. When the enhancin-like mutant was fed to Galleria mellonella larvae, no significant reduction in virulence was observed.In the present study, we report a B. thuringiensis enhancin-like gene (bel) encoding a protein (Bel) that has 20 to 30% identity to the viral enhancin protein and 95% identity to bacterial enhancin-like proteins. Therefore, Bel function may have a synergistic action similar to that of the virus enhancin protein. To understand the biochemical activity of this novel bacterial gene, bel was knocked out in the plasmid-free strain BMB171. We expected that this bel mutant would have no significant reduction in toxicity according to the reports of Galloway et al. (8) and Hajaij-Ellouze et al. (12). However, the bel mutant surprisingly resulted in dramatically reduced Cry1Ac toxicity to Helicoverpa armigera larvae. To further confirm this result, purified Bel was fed together with the Cry1Ac protein to H. armigera larvae. We found that Bel can function as a synergist of Cry1Ac toxicity against H. armigera. In vivo and in vitro observations showed that Bel can disrupt the insect midgut PM and degrade IIM of insect midgut PM. The target of Bel is the IIM of PM, similar to the results found with viral enhancin.     

Susceptibility to Cry proteins of a Spanish Ostrinia nubilalis glasshouse population repeatedly sprayed with Bacillus thuringiensis formulations

Ostrinia nubilalis Hübner (Lepidoptera: Crambidae), a major pest of corn in temperate climates, can feed on other crops due to its polyphagous behaviour. In particular, this species became a serious problem in some sweet pepper commercial glasshouses in south‐eastern Spain repeatedly sprayed with Bacillus thuringiensis (Bt) products to control Spodoptera exigua Hübner (Lepidoptera: Noctuidae). The susceptibility of an O. nubilalis colony established from individuals collected in these Bt‐sprayed glasshouses was compared with a reference laboratory colony. Differences in susceptibility between the two colonies to Cry1Aa, Cry1Ab, Cry1Ac and Cry2Aa proteins were found. However, our results indicate that the O. nubilalis control failure in the glasshouse was not due to selection for resistance. Intraspecific variation probably accounts for differences between the glasshouse‐derived population and the laboratory strain. This conclusion is based on several lines of evidence: the glasshouse‐derived population retained its susceptibility to a Bt standard product and to most of its individual components (both in the form of protoxins and in the form of activated toxins), and it did not respond to laboratory selection with high doses of Cry1Ab.     

Midgut cysteine protease-inhibiting activity in Trichoplusia ni protects the peritrophic membrane from degradation by plant cysteine proteases

The action of plant cysteine proteases on the midgut peritrophic membrane (PM) of a polyphagous herbivorous lepidopteran, Trichoplusia ni, was studied. Proteins in PMs isolated from T. ni larvae were confirmed to be highly resistant to the serine proteinases trypsin and chymotrypsin, but were susceptible to degradation by plant cysteine proteases, which is consistent with the known molecular and biochemical characteristics of the T. ni PM proteins. However, the PM proteins were not degraded by plant cysteine proteases in larvae or in the presence of larval midgut fluid in vitro. With further biochemical analysis, cysteine protease-inhibiting activity was identified in the midgut fluid of T. ni larvae. The cysteine protease-inhibiting activity was heat resistant and active in the tested pH range from 6.0 to 10.0, but could be suppressed by thiol reducing reagents or reduced by treatment with catalase. In addition to T. ni, cysteine protease-inhibiting activity was also identified from two other polyphagous Lepidoptera species, Helicoverpa zea and Heliothis virescens. In conclusion, results from this study uncovered that herbivorous insects may counteract the attack of plant cysteine proteases on the PM by inhibiting the potentially insecticidal cysteine proteases from plants in the digestive tract. However, the biochemical identity of the cysteine protease-inhibiting activity in midgut fluid has yet to be identified.     

The evaluation of anti juvenile hormones using last stadium larvae of the cabbage looper,Trichoplusia ni (Hübner)

Treatment of prepupae of the cabbage looper, Trichoplusia ni (Hübner), with the anti juvenile hormone (AJH) fluoromevalonolactone (FMev) reduces juvenile hormone esterase (JHE) activity and either delays or inhibits the normal larval-pupal ecdysis in a dose-dependent manner, resulting in the formation of pupae with a suite of characteristic abnormalities. A variety of putative AJHs were evaluated using these teratogenic effects and changes in JHE activity as indicators of AJH activity. Like FMev, the compactin analog L-643,049-01K01 and, at very high doses, 3,3-dichloro-2-propenyl hexanoate (DPH) behaved as AJHs. FMev and L-643,049-01K01 were most effective when larvae were treated at gut purge, while DPH was the most effective after the initiation of spinning behavior.     

Tomato variety affects larval survival but not female preference of the generalist moth Trichoplusia ni

In herbivorous insects, the choice that females make for a suitable host plant is crucial for survival of its offspring because the neonate larvae are generally not capable of moving great distances. The preference-performance hypothesis states that herbivore females will choose to oviposit on hosts on which their offspring will have better performance. In this study, we investigated whether Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae) females are able to discriminate among a weedy race, a landrace, and a commercial cultivar of tomato plants, Solanum lycopersicum L. (Solanaceae), and how their choice affects the offspring performance. Additionally, we identified the volatile compounds and recorded the density of glandular trichomes of the tomato plants. Females did not show a preference for any of the three types of tomato plants. Females oviposited more on the adaxial surface of leaves of commercial cultivar plants than on (any surface of) leaves of weedy-race plants. The relative abundance of volatiles varied quantitatively among the three types of tomato plants. Commercial cultivar plants released a higher abundance of volatiles than weedy race and landrace plants. Weedy-race plants had a higher density of glandular trichomes types I and VI than the commercial cultivar. More neonate larvae died if fed on the weedy race and landrace plants than when reared on commercial cultivar plants. Results suggested that the higher mortality of T. ni larvae may be related to a higher density of glandular trichomes on landrace and weedy-race plants than on commercial cultivar plants, although other constitutive and induced defenses may be involved. Our results do not support the preference-performance hypothesis.     

A baculovirus enhancin alters the permeability of a mucosal midgut peritrophic matrix from lepidopteran larvae

The peritrophic matrix (PM) in lepidopterous larvae may function as a defensive barrier against ingested viral pathogens. PMs isolated from Trichoplusia ni and Pseudaletia unipuncta larvae, were treated with a baculovirus-encoded metalloprotease (enhancin) from Trichoplusia ni granulosis virus (TnGV) and their in vitro permeability to blue dextran and fluorescent-labelled Autographa californica nuclear polyhedrosis virus (AcMNPV) was determined using a dual chamber permeability apparatus. Incubation of T. ni PMs with 0.0, 0.5, 1.0, and 2.0mg/ml enhancin resulted in a blue dextran 2000 flux of 4.4, 6.3, 9.9, and 15.6&mgr;g/mm(2)/h, respectively. In addition, T. ni PMs treated with enhancin were found to be significantly more permeable to fluorescent-labelled AcMNPV than non-treated control PMs. The permeability of T. ni PMs treated with 3.0mg/ml enhancin was 0.017 cumulative percent crossing/mm(2)/h, whereas the permeability of the control PM was below the detectable limit. Similarly, enhancin treatment greatly increased the permeability of P. unipuncta PMs to AcMNPV. These results provide evidence that the PM from two lepidopteran species can block the passage of baculovirions across this matrix thus reducing the probability of larval infection. Furthermore, these results support the hypothesis that enhancin facilitates NPV infection of larvae by altering the permeability of the PM.     

Stay or move: how Bt‐susceptible Helicoverpa armigera neonates behave on Bt cotton plants

Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) larvae occasionally have been reported to survive at management threshold levels in fields of Bollgard II^® cotton, Gossypium hirsutum L. (Malvaceae). The pattern and degree of larval survival is not easily predicted but depends on the ability of first instars to establish on host plants. Experiments were conducted with Bacillus thuringiensis Berliner (Bt)‐susceptible and Bt‐resistant larvae of H. armigera to understand how physiologically Bt‐susceptible H. armigera survive on Bt cotton plants, and examine how their first meal influences survival rates. In assays using cotton plant parts, both strains of larvae displayed similar tendencies to drop‐off specific plant parts of Bt and non‐Bt cotton. However, significantly more Bt‐susceptible larvae dropped off young leaves, mature leaves, and squares of Bt cotton compared to non‐Bt cotton plants. Egg cannibalism significantly improved the survival of Bt‐susceptible H. armigera larvae on Bt cotton plants. Larvae were more likely to eat live aged eggs, than newly laid or dead eggs. Survival significantly improved when larvae cannibalized eggs before feeding on Bt leaves. The behavior of Bt‐susceptible larvae with respect to drop‐off and egg cannibalism may help enhance their survival on Bt cotton plants.     

粘虫颗粒体病毒增效蛋白基因片段优化及功能

【目的】研究转宿主粘虫颗粒体病毒(Pseudaletia unipuncta granulovirus,Pu GV-Ps)增效蛋白基因截短片段优化及其增效作用,探索增效蛋白基因的合理利用途径。【方法】生物信息学分析增效蛋白结构域,构建增效蛋白基因截短片段原核表达载体,分析目的基因片段表达产物的表达水平、围食膜蛋白降解效能和增强活性,进一步明确Pu GV-Ps增效蛋白基因的功能区域。【结果】Pu GV-Ps增效蛋白含有M60-like结构域、锌离子催化域和糖蛋白结合域,并包含13个潜在的糖基化位点。以此为依据设计P69(短截M60-like结构域)和P77(短截糖蛋白结合域)2个截短片段,构建了表达载体p ET15b-P69和p ET15bP77,原核表达量明显高于全长基因P104。表达产物纯化蛋白围食膜降解活性表明,P69对斜纹夜蛾围食膜大分子蛋白降解程度高于P77,但两者均低于P104。病毒增强苏云金杆菌(Bt)实验表明,截短片段的表达产物提高了Bt对小菜蛾的毒力,但增强活性显著低于P104。【结论】研究结果表明,Pu GV-Ps增效蛋白基因N端M60-like结构域和C端糖蛋白结合域对其增效作用的发挥都具有一定功能,这些结构对维持增效蛋白的构象也发挥了一定的作用,截短片段P69有利于保持Pu GV-Ps增效蛋白的活性、提高表达水平。该研究结果对增效蛋白的工业化生产具有一定的指导意义。     

寄主植物对棉铃虫越冬蛹抗寒能力的影响

通过对滞育蛹过冷却点的测定 ,初步明确寄主对棉铃虫Helicoverpaarmigera的越冬抗寒性有影响 ,幼虫取食棉花时产生的滞育蛹的过冷却点低于取食玉米的滞育蛹的过冷却点 ,即前者的抗寒能力高于后者 ;但用Bt棉喂养后的棉铃虫滞育蛹抗寒能力下降